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Analyse protéomique ciblée à haute résolution : un outil puissant pour le diagnostic clinique à partir de prélèvements de tissus amyloïdes bruts

Abstract : Amyloidosis is rare disease that appears as an insoluble deposition of specific extracellular proteins. Thirty classes of amyloidosis have been reported and their diagnosis relies on the identification of the associated proteins by immunohistochemistry analysis of the affected tissues. However, its interpretation is highly dependent on the pathologist and can be ambiguous. Laser capture micro-dissection (LCM) and mass spectrometry are a promising alternative. We have developed a targeted proteomics approach from raw tissues treated with ultrasound. We showed that the biopsies taken from patients presenting different classes of amyloidosis can be proteolysed by ultrasonic treatment. This technique increases the efficiency and reduces the time of digestion (90s instead of 15h), while minimizing the amount of material needed. After a first unbiased proteomics study for clinical diagnosis of patients with amyloidosis in various organs/tissues (kidney, lung, salivary glands, testicle, vitreous humor and spleen), we have optimized a dedicated screening method for a technological transfer to the French clinical departments. For this, two methods show very promising results: SRM (selected reaction monitoring) on a low-resolution mass spectrometer, available in some clinical laboratories, and PRM (parallel reaction monitoring) available on the latest generation of high-resolution instruments which provide fast results. In conclusion, we propose here a specific protocol for a robust diagnosis and rapid classification of amyloidoses.
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Sophie Liuu. Analyse protéomique ciblée à haute résolution : un outil puissant pour le diagnostic clinique à partir de prélèvements de tissus amyloïdes bruts. Chimie-Physique [physics.chem-ph]. Université Pierre et Marie Curie - Paris VI, 2014. Français. ⟨NNT : 2014PA066373⟩. ⟨tel-01403464⟩

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